CATALYTIC COMPETENCE AND REGULATION OF RECEPTOR KINASE

受体激酶的催化能力和调节

• 批准号: 3233521
• 负责人: JULES Alan SHAFER
• 金额: $ 10.15万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 1985
• 资助国家: 美国
• 起止时间: 1985-08-01 至 1989-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3233521
• 关键词: catalyst; chemical reaction; conformation; enzyme substrate; gel electrophoresis; glucose transport; high performance liquid chromatography; human tissue; immunochemistry; phosphorylation; protein tyrosine kinase; stoichiometry; zinc

The intact phosphorylated and unphosphorylated forms of insulin receptor kinase will be purified from human placenta by chromatography on immobilized anti-O-phosphotyrosyl antibody. The purified forms of the receptor will be used to characterize the multistep reaction pathway for the conversion of the unphosphorylated form of the receptor to the catalytically active phosphorylated form. In these studies the amino acid sequence around the reactive tyrosyl residue(s) in the receptor will be determined and compared to that of other tyrosine kinases. Rate and equilibrium constants will be evaluated for individual steps in the reaction pathway for receptor activation. Stoichiometry and equilibirum constants for the binding of insulin to the phosphorylated and unphosphorylated forms of the receptor will be measured to determine whether insulin binding might cause the unphosphorylated receptor to assume a conformation similar to that of the phosphorylated receptor. The inhibitory effect of zinc ion on individual steps in the reaction pathway for receptor activation will be characterized, since inhibition of autophosphorylation of the insulin receptor by zinc ion might serve to protect the islet cells and other tissues from deleterious effects of high concentrations of insulin at the site of normal or accidental release of the contents of insulin-containing granules. Another set of studies will be directed toward characterization of interactions of the receptor kinase with potential regulators of metabolism. Thus, the possibility will be assessed that insulin receptor kinase catalyzes phosphorylation of the glucose transporter protein and thereby decreases its exit rate from plasma membranes. The phosphorylation of insulin receptor by casein kinase I will be studied to determine whether casein kinase I catalyzed phosphorylation of insulin receptor alters its ability to bind insulin and undergo autophosphorylation, and whether casein kinase I catalyzed phosphorylations of the receptor might account for the phosphorylation at receptor seryl and thereonyl residues seen in whole cells. In a search for endogenous substrates for the insulin receptor kinase, anti-O-phosphotyrosyl antibody will be used to concentrate products of insulin promoted tyrosyl phosphorylations in whole cells. In an attempt to deduce the structural characteristics of natural substrates for the insulin receptor kinase, the substrate specificity of this enzyme toward nonpeptide and peptide substrates will be determined.

胰岛素受体完整的磷酸化和未磷酸化形式

项目成果

Phosphorylation of highly purified growth hormone receptors by a growth hormone receptor-associated tyrosine kinase.

生长激素受体相关酪氨酸激酶对高度纯化的生长激素受体进行磷酸化。

• 发表时间: 1989
• 期刊: The Journal of biological chemistry
• 作者: Carter-Su,C;Stubbart,JR;Wang,XY;Stred,SE;Argetsinger,LS;Shafer,JA
• 通讯作者: Shafer,JA

Growth hormone promoted tyrosyl phosphorylation of growth hormone receptors in murine 3T3-F442A fibroblasts and adipocytes.

生长激素促进小鼠 3T3-F442A 成纤维细胞和脂肪细胞中生长激素受体的酪氨酰磷酸化。

• DOI: 10.1021/bi00401a049
• 发表时间: 1988
• 期刊: Biochemistry
• 影响因子: 2.9
• 作者: Foster,CM;Shafer,JA;Rozsa,FW;Wang,XY;Lewis,SD;Renken,DA;Natale,JE;Schwartz,J;Carter-Su,C
• 通讯作者: Carter-Su,C